Preparation Of Monoclonal Antibody Against Chloramphenicol And Establishment Of Immunoassay Method For Salbutamol

Chloramphenicol(CAP), also known as L-chloramhenicol, L-neomycin, Chloraminebenzene alcohol, it is a kind of relatively inexpensive and highly effective broad-spectrumantibacterial antibiotics. CAP is toxic, and it has the toxic side efficts to the human beings andanimals by using it, for examples it can make the female animals abort, preterm，make theanimals lower fertility, reduce the fecundity and so on. CAP can also give the human beingsmany disease, such as optic neuritis, gray syndrome, liver injury, blood disease, CAP can evenlead to the death.At present, the detection technologies of CAP are mainly Microbiological method,ELISA, HPLC, GC-MS, LC-MS and so on. Microbiological method has the poor specificity,low sensitivity, needs more time. HPLC and GC-MS can detect the antibiotic on the low levelof the remains, and the specifity is strong. But the instruments are very expensive, the time ofadvance process of samples are too long. ELISA has the features of high sensitivity, strongspecificity, short time, and also it can detect many samples the same time, it doesn’t need theexpensive instruments, it suits the primary laboratory personnel determining lots of examines.This experiment is examing the CAP remains by using ELISA.This experiment uses the valuing nitrogen method aiming at the CAPC’s nitro, choosesBSA and OVA as carrier protein, couples the CAP’s micromolecule. When the couple is done,we use the SDS-PAGE electro-pulse method, UV-light spectrum method, infrared scanningatlas method’s prove that CAP whole antigen compound experiment is successful, and therates are: CAP-BSA17.1:1,CAP-OVA10:1.We use CAP-BSA as the immunogen letting three Balb/c mice immune,and use theindirect ELISA method to examine the serum’s antibody titer, the antibody titers separatelyare1：6400,1：6400,1：12800.We also examine the mice blood’s sensibility, and choose theNO.3mouse to do the fusion experiment because its blood’s titer and sensibility are the best.After the cell fusion experiment, we choose three hybrid tumor cells which has good effectshybridoma, they are separately2A6,4E10and4E12. We inject these three cells seperatelyinto the Balb/c mice’s abdominal cavity, after7days, we select the abdominal cavity water andexamine the titers, and2A6、4E10、4E12’s hybrid tumor cells’ clear solution titers are:1： 12800、1:6400、1:6400, the abdominal cavity water’s antibody titers are1:51200、1:25600、1:25600. Next, we choose2A6’s antibody, and use the ELISA method to examine itssensitivity and specificity. The rusults shows that the2A6’s antibody is aiming at CAP whichhas good specificity, and also the CAP standard samples‘IC50number is3.125ppb. We alsomake cross reaction experiments on CAP’s analoge, and the cross reaction rates betweenCAP’s2A6antibody and florfeni, propacin, colTCs, sulfadimidine, it also prove that antibodyCAP’s monoclonal antibody has the good specificity.This experiment uses indirect competition ELISA method to detect CAP’s remains, andthe rusults show that the sensitivity can reach2.213ppb, but the standard samples’ sensitivityis only0.17ppb. This study makes CAP’s monoclonal antibody, establishes at a preliminarylevel a ELISA method to PLA, otimizes the Antigen and Antibody’s reaction time, firstantiviral concentration, antigen coated conditions, ELIAS secondary antibody concentrationand peridium concentration which are in the reaction condition. This experiment providestheoretical basis for further studying detection samples of PLA’s remains.